The neurotoxins of Clostridium botulinum (BoNTs) are the most toxic proteins to humans and are Category A agents. BoNTs (approximately 150,000 kDa) are zinc-dependent proteases that comprise a group of seven proteins designated as serotypes A through G. BoNT(A-G) share similar AB structure-function properties, but possess limited primary amino acid homology. BoNTs elicit flaccid paralysis through the cleavage of SNARE proteins that are part of the trafficking and docking components of fusion vesicles. The B component of BoNT binds to receptors located on neuronal cells, yielding the trophism of this intoxication. While the paralysis elicited by the A-G serotypes of BoNT is clinically similar, each serotype of BoNT cleaves individual SNARE proteins at specific amino acids. NIAID has identified the development of therapeutics and vaccines against BoNT as high priority for bio-defense products. This core facility will support the research efforts of two research proposals within Region 5: "Vaccines and therapeutics against botulism" and "Nanofluidic Transduction of Stochastic Biomolecular Recognition of BoNT". While investigators involved in these proposals have expertise in specific aspects of bacterial pathogenesis, Dr. Johnson provides expertise in BoNT biology and genetics. The core facility will: Maintain strains of Clostridium botulinum and nontoxigenic derivatives of C. botulinum for genetic studies; Produce and purify BoNTs; Provide expertise on stabilization of BoNTs to retain high specific toxicity; Provide BoNTs and genetic materials from C. botulinum and nontoxigenic derivatives of C. botulinum to members of the RCE; Conduct genetic conjugation/transformation and recombinant DNA manipulations on C. botulinum; Produce and purify mutated forms of BoNT in C. botulinum; Conduct animal studies, including mouse LD50s, mouse challenges, and neutralization analyses; and maintain biosafety environment for research with C. botulinum and BoNTs. Developing this core will provide the platform for current research efforts of the RCE and facilitate future projects related to BoNT research within Region 5.